Abdul Halim Harrath

Evaluation of cytotoxic, oxidative stress, proinflammatory and genotoxic effect of silver nanoparticles in human lung epithelial cells.

Silver nanoparticles are increasingly used in various products, due to their antibacterial properties. Despite its wide spread use, only little information on possible adverse health effects exists. Therefore, the aim of this study was to assess the toxic potential of silver nanoparticles (<100 nm) in human lung epithelial (A549) cells and the underlying mechanism of its cellular toxicity. Silver nanoparticles induced dose and time‐dependent cytotoxicity in A549 cells demonstrated by MTT and LDH assays. Silver nanoparticles were also found to induce oxidative stress in dose and time‐dependent manner indicated by depletion of GSH and induction of ROS, LPO, SOD, and catalase. Further, the activities of caspases and the level of proinflammatory cytokines, namely interleukin‐1β (IL‐1β) and interleukin‐6 (IL‐6) were significantly higher in treated cells. DNA damage, as measured by single cell gel electrophoresis, was also dose and time‐dependent signicants in A549 cells. This study investigating the effects of silver nanoparticles in human lung epithelial cells has provided valuable insights into the mechanism of potential toxicity induced by silver nanoparticles and warrants more careful assessment of silver nanoparticles before their industrial applications.

The breadth of the placental surface but not the length is associated with body size at birth

Studies of pregnancies complicated by preeclampsia led to the suggestion that the surface of the placenta is aligned along two axes, measured by its breadth and length. It was hypothesised that tissue along the breadth serves as a nutrient sensor, responding to the mothers nutritional state and fetal nutritional demands, while tissue along the length has different functions. To develop this hypothesis we measured the breadth and length of the placental surface in 401 neonates born in the King Khalid Hospital, Riyadh, Saudi Arabia, and related these measurements to the babys body size. The breadth and length of the placental surface were highly correlated (coefficient = 0.7). Nevertheless, in a simultaneous regression with both measurements, only the breadth was associated with neonatal body size. There were strong trends of increasing birth weight, ponderal index, and the circumferences of the head, chest, abdomen and thigh with increasing placental breadth. In contrast no measurement of babys body size was related to placental length. Birth weight increased by 125 g per cm increase in placental breadth (95% confidence interval 88 to 162, p < 0.001) but only by 20 g per cm increase in placental length (-13 to 53, p = 0.2). The corresponding figures for head circumference were 0.28 cm (0.17-0.39, p < 0.001) and 0.03 (-0.07 to 0.14, p = 0.5). The associations between placental breadth and neonatal body size were strongest if the mothers height was below the median (157 cm). The associations between a larger breadth of the placental surface and a larger baby are consistent with the hypothesis that tissue along the breadth plays a key role in nutrient transfer from mother to baby. Mothers who are short in stature are known to have lower rates of protein turnover in pregnancy. In these circumstances the ability of the placenta to transfer amino acids to the fetus may be critical.

Schmidtea mediterranea phylogeography: an old species surviving on a few Mediterranean islands?

BackgroundSchmidtea mediterranea (Platyhelminthes, Tricladida, Continenticola) is found in scattered localities on a few islands and in coastal areas of the western Mediterranean. Although S. mediterranea is the object of many regeneration studies, little is known about its evolutionary history. Its present distribution has been proposed to stem from the fragmentation and migration of the Corsica-Sardinia microplate during the formation of the western Mediterranean basin, which implies an ancient origin for the species. To test this hypothesis, we obtained a large number of samples from across its distribution area. Using known and new molecular markers and, for the first time in planarians, a molecular clock, we analysed the genetic variability and demographic parameters within the species and between its sexual and asexual populations to estimate when they diverged.ResultsA total of 2 kb from three markers (COI, CYB and a nuclear intron N13) was amplified from ~200 specimens. Molecular data clustered the studied populations into three groups that correspond to the west, central and southeastern geographical locations of the current distribution of S. mediterranea. Mitochondrial genes show low haplotype and nucleotide diversity within populations but demonstrate higher values when all individuals are considered. The nuclear marker shows higher values of genetic diversity than the mitochondrial genes at the population level, but asexual populations present lower variability than the sexual ones. Neutrality tests are significant for some populations. Phylogenetic and dating analyses show the three groups to be monophyletic, with the west group being the basal group. The time when the diversification of the species occurred is between ~20 and ~4 mya, although the asexual nature of the western populations could have affected the dating analyses.ConclusionsS. mediterranea is an old species that is sparsely distributed in a harsh habitat, which is probably the consequence of the migration of the Corsica-Sardinia block. This species probably adapted to temperate climates in the middle of a changing Mediterranean climate that eventually became dry and hot. These data also suggest that in the mainland localities of Europe and Africa, sexual individuals of S. mediterranea are being replaced by asexual individuals that are either conspecific or are from other species that are better adapted to the Mediterranean climate.

The involvement of SIRT1 and transcription factor NF-κB (p50/p65) in regulation of porcine ovarian cell function

The role of either mTOR system/enzyme sirtuin1 (SIRT1) or transcription factor NF-κB in the direct control of ovarian function has not been estabished. The aim of our in vitro experiments was to examine the involvement of SIRT1 and the p65 and p50 subunits of NFκB in control of porcine ovarian granulosa cell functions and the interrelationships between SIRT1, NFκB (p65, p50) 30 and FSH in the ovary. Monolayers of primary granulosa cells were transfected with gene constructs encoding either SIRT1 or p65 and p50, and thereafter cultured with, or without, addition of FSH. The accumulation of markers of proliferation (cyclin B1 and cyclin-dependent protein kinase Cdc2/p34) and proteins p50, p65 and SIRT1 in the cells was detected by using SDS-PAGE/Western immunoblotting and immunocytochemistry. The secretion of progesterone (P4) and insulin-like growth factor I (IGF-I) was measured by using radioimmunoassay. It was observed that transfection of cells with a SIRT1 gene construct promoted accumulation of proliferation markers, Cdc2/p34, cyclin B1, decreased accumulation of p50 and p65 and stimulated release of P4 and IGF-I. Co-transfection of cells with cDNA p50 and cDNA p65 enhanced the accumulation of SIRT1 and the release of P4 but did not influence the release of IGF-I. Adding FSH to the culture medium stimulated accumulation of both subunits of NF-κB, as well as accumulation of Cdc2/p34, cyclin B1 and release of both P4 and IGF-I. The ability of FSH to promote NF-κB accumulation, the similarity of the main effects of FSH, SIRT1 and NF-κB, as well as the inability of NF-κB to substantially modify the the majority of FSH effects suggest that SIRT1/NF-κB system could be a mediator of FSH action on ovarian cell functions. On the other hand, SIRT1 was able to inhibit NF-κB and to change stimulatory the effect of FSH on NF-κB from stimulatory to inhibitory. This could suggest the existence of negative feedback control of FSH/NF-κB system by high amounts of SIRT1. Our observations (1) confirm the previous data on proliferation, P4 and IGF-I release in ovarian cells and their up-regulation by FSH, (2) demonstrate the presence of SIRT1, NF-κB/p50 and NF-κB/p65 in these cells, (3) show for the first time the involvement of SIRT1 and NF-κB in direct control of proliferation and secretory activity of ovarian cells, (4) represent the first data on interrelationships between FSH, SIRT1 and NF-κB within the ovary.

Intergenerational effects of in utero exposure to Ramadan in Tunisia

We have reported that changes in the lifestyle of pregnant women during Ramadan affect more than one generation. In a series of newborn babies in Saudi Arabia, those whose mothers had been in utero during Ramadan differed from those whose mothers had not been in utero during Ramadan. These were unexpected findings and require replication.

Sensitivity of freshwater pulmonate snail Lymnaea luteola L., to silver nanoparticles

Toxicity of nanoparticles depends on many factors including size, shape, chemical composition, surface area and surface charge. Silver nanoparticles (AgNPs) are likely to enter the aquatic ecosystems because of their multiple applications and pose a health concern for humans and aquatic species. Therefore, we used a freshwater snail Lymnaea luteola L (L. luteola) to investigate the acute toxicity and genotoxicity of AgNPs in a static-renewal system for 96 h. AgNPs caused molluscicidal activity in L. luteola, with 96-h median lethal concentrations (LC50) (48.10 μg L(-1)). We have observed that AgNPs (36 μg L(-1)) elicited a significant (p<0.01) reduction in glutathione, glutathione-s-transferase and glutathione peroxidase with a concomitant increase in malondialdehyde level and catalase in digestive gland of L. luteola. However, a significant (p<0.01) induction in DNA damage was observed by the alkaline single cell gel electrophoresis in digestive gland cells treated with AgNPs for 24 and 96 h. These results demonstrate that silver nanoparticles are lethal to freshwater snail L. luteola. The oxidative stress biomarkers and comet assay can successfully be used as sensitive tools of aquatic pollution biomonitoring.

First report of adelphophagy in flatworms during the embryonic development of the planarian Schmidtea mediterranea (Benazzi, Baguñà, Ballester, Puccinelli & Del Papa, 1975) (Platyhelminthes, Tricladida)

Abstract This paper details the embryonic development of the triclad flatworm Schmidtea mediterranea and describes, for the first time in flatworms, the phenomenon of adelphophagy. Embryonic development, which takes 23 days at 18–20°C, basically corresponds with the developmental stages documented in other studies for triclad flatworms. At some of the stages it was observed occasionally that some embryos cannibalized their siblings, i.e. exhibited adelphophagy or embryophagy. It is suggested that this phenomenon may contribute to the fact that the number of young hatching from a cocoon is usually considerably less than the number of egg cells initially assembled in the capsule. The occurrence of adelphophagy in the animal kingdom is discussed.

Étude ultrastructurale de l’ovogenèse chez la planaire Schmidtea mediterranea (Plathelminthe, Paludicole)

The ovary of the freshwater planarian Schmidtea mediterranea has been studied for the first time using both light and electron microscopy methods. The ultrastructure of the ovary revealed two types of cells: accessory cells and germinal cells at various stages of differentiation, distributed along a maturation axis. Initially, oogonia underwent cytoplasm growth due to the development of organelles, such as endoplasmic reticulum, Golgi complex, and mitochondria, which are all involved in the production of cytoplasmic inclusions or yolk globules. It is shown that the chromatoid body and fibrogranular aggregates may participate in the synthesis of vitelline inclusions. When completely mature, the oocytes have become larger, due to the accumulation of nutritive inclusions, which are round in shape and have a paracrystalline structure. These inclusions are interpreted as being yolk globules and may represent a kind of nutritive material for the developing embryo. These ultrastructural features of the ovary agree with the available phylogenetic tree, based on morphological and karyological characters that considers Schmidtea group as a genus and not a subgenus. The presence of sperm between the oocytes suggests that fertilization may occur within the ovary, representing an uncommon condition within the Triclads, in which fertilization usually takes places outside of the ovaries.

Reproductive Strategies, Karyology, Parasites, and Taxonomic Status of Dugesia Populations from Yemen (Platyhelminthes: Tricladida: Dugesiidae)

We present new data on the distribution, reproductive strategies, karyology, and taxonomic status of populations of freshwater planarians from Yemen. Nine populations were sampled and significant differences in their reproductive strategies and karyology are reported. The present study presents the first fully documented record of a naturally sexual, diploid (2n = 18) population of a Dugesia species in the eastern part of the Afrotropical region. Morphological characters combined with karyological data suggest that these Dugesia populations from Yemen represent a new species, which is herein described as Dugesia arabica Harrath and Sluys, sp. nov. This new species is mainly distinguishable from other Dugesia species that are distributed exclusively in the Mediterranean basin and in the eastern part of the Afrotropical region by the presence of the following features: well-developed and cone-shaped penis papilla, housing an ejaculatory duct that runs ventrally and has a subterminal and ventral opening; a considerably expanded and folded section of the bursal canal at the level of the oviducal openings; absence of a layer of longitudinal muscles on the copulatory bursa and the bursal canal. Specimens from two populations from Yemen were infested with a gregarine Protozoon.

THE FIRST SUBTERRANEAN FRESHWATER PLANARIANS FROM NORTH AFRICA, WITH AN ANALYSIS OF ADENODACTYL STRUCTURE IN THE GENUS DENDROCOELUM (PLATYHELMINTHES, TRICLADIDA, DENDROCOELIDAE)

The paper describes the first species of freshwater planarians collected from subterranean localities in northern Africa, represented by three new species of Dendrocoelum Orsted, 1844 from Tunisian springs. Each of the new species possesses a well-developed adenodactyl, resembling similar structures in other species of Dendrocoelum, notably those from southeastern Europe. Comparative studies revealed previously unreported details and variability in the anatomy of these structures, particularly in the composition of the musculature. An account of this variability is provided, and it is argued that the anatomical structure of adenodactyls may provide useful taxonomic information.